Microsatellite DNA reveals genetically different populations of Atlantic bonito Sarda sarda in the Mediterranean Basin

Microsatellites were used to investigate population genetic structure of Atlantic bonito Sarda sarda from the Black Sea, Marmara Sea, Aegean Sea, north-eastern Mediterranean Sea and Adriatic Sea. Overall average observed heterozygosity was high (0.93). Average observed heterozygosity per locus ranged from 0.79 to 0.98. Pairwise F_ST estimates for all loci between populations ranged from 0 to 0.07626, and significant F_ST values (P < 0.001) were detected between populations; the Blacks Sea and Marmara Sea samples were not significantly different from each other, but significant different from the other samples, and Aegean Sea and north-eastern Mediterranean Sea samples were also not significantly different from each other, but significantly different from all other samples. The Adriatic Sea sample was significant different from all other samples. The Mantel test revealed a significant (P < 0.001, r = 0.68) isolation-by-distance for these 11 populations. Neighbour-joining analysis clustered the Black Sea and Marmara Sea samples together while collections from Aegean Sea and north-eastern Mediterranean Sea were clustered close to each other and far from the others. On the other hand, the Adriatic Sea collection presented very distinctive relationship from the others.     

Diet of the Atlantic bonito Sarda sarda (Bloch, 1793) in the Northeast Aegean Sea

Seasonal and size‐related patterns in feeding habits of the Atlantic bonito (Sarda sarda) in the Northeast Aegean were investigated over a 2‐year period May 2007 – May 2009 based on analysis of stomach contents of 134 fish ranging from 266–655 mm fork length. The fish were caught by purse‐seine boats operating around the islands of Lesvos and Chios (Greece). Of the stomachs examined, 46% contained at least one prey item and significant seasonal and size‐related patterns were found in the occurrence of non‐empty stomachs. The overall diet composition in terms of numerical importance was 71% fish, 28% crustaceans and 1% cephalopods. Of the fish, Engraulidae (Engraulis encrasicolus) were the dominant prey taxon, followed by Centracanthidae (Spicara spp.) and then Clupeidae (Sardina pilchardus). Statistically significant monthly variation in diet was apparent, suggesting that S. sarda take advantage of seasonally abundant prey fish in the study area, but no size‐related variation in the importance of the main prey species was detected.     

Biochemical composition of the Atlantic bonito Sarda sarda from the Aegean Sea (eastern Mediterranean Sea) in different stages of sexual maturity

The content (% wet mass) in water, ash, lipid, crude protein, DNA and RNA of different tissues was determined during sexual maturation of bonitos Sarda sarda from the Aegean Sea. A total of 220 specimens were collected in the following stages of sexual maturity: immature, resting, developing, mature, spawning and spent. Highest lipid levels in the white muscle, red muscle and liver were measured in immature specimens, while lowest levels were found in spawning bonitos. The gradual percentage of lipid reduction from immature to spawning bonitos was relatively higher in the liver (females 71·2% and males 64·4%) than in the white (females 59·2% and males 53·5%) and red (females 62·1% and males 51·7%) muscle. Lipid levels in the gonads increased gradually from the immature to spawning stage. The decrease of lipid in the somatic tissues was more intense in females than in males, and gonadal lipid content was higher in females than in males. There was a strong reverse correlation between water and lipid percentage in all tissues. Protein content decreased significantly only in spawning bonitos. The percentage of protein reduction from immature to spawning stage was relatively higher in males than in females in both white (females 3·4% and males 4·6%) and red (females 4·6% and males 5·1%) muscles. Protein content in the liver was significantly lower than in the other tissues, being highest in mature females. Gonadal protein content in females increased with maturation and decreased after spawning. The content in ash exhibited considerable stability. The RNA:DNA ratio exhibited a similar pattern of variation in both muscles. The RNA:DNA ratio increased during gonadal development gradually from the developing to spent stage. It was concluded that in S. sarda during gonadal development, there was an increase in gonadal lipid accompanied by a decrease in somatic tissue lipid reserves. Thus, reproductive inactive bonitos have more lipid in their edible part and a higher nutritional value than active ones.     

Age and growth of Atlantic bonito (Sarda sarda Bloch, 1793) in the Sea of Marmara and Black Sea,Turkey

Age and growth of Atlantic bonito (Sarda sarda Bloch, 1793) were determined in the Black Sea and Sea of Marmara from October 2003 to May 2005. A total of 694 samples ranging from 23.5 to 71 cm total length was collected; age estimates were carried out on otoliths of 248 fish. Male and female fish ages were estimated as up to 3⁺. The length–weight relationship and the von Bertalanffy growth curve (TVBGF) were described for all individuals as: W = 0.0054L^3.215 and L_t = 68.01 [1 ? e^{?0.895 (t + 0.395)}]. These data are discussed in relation to other growth studies of S. Sarda in the Mediterranean Sea.     

Phylogeography of Xiphorhynchus fuscus (Passeriformes, Dendrocolaptidae): vicariance and recent demographic expansion in southern Atlantic forest

Knowledge of the evolutionary processes that shaped a biota is important for both academic and conservation purposes. The objective of the present study is to analyse the mitochondrial genetic variation of Xiphorhynchus fuscus (Aves: Dendrocolaptidae) from the southern Atlantic forest in Brazil and Argentina, and to discuss whether the results support different hypotheses regarding the local intraspecific diversification of this species . We sequenced 575 bp of the control region of 114 specimens collected in the Brazilian states of Bahia, Minas Gerais, Rio de Janeiro, São Paulo, Paraná, and Santa Catarina, and in the province of Misiones in Argentina. We studied the population genetic structure with analysis of molecular variance and the demographic history with multiple regression analysis, coalescence simulations, and demographic tests. Xiphorhynchus fuscus presented a significant population genetic structure (Φ_st = 0.57). Three mitochondrial lineages were described, one associated with Xiphorhynchus fuscus tenuirostris and the others with Xiphorhynchus fuscus fuscus. The data did not support the primary influence of geographical barriers or rivers in the intraspecific diversification of X. fuscus in the southern Atlantic forest. Instead, the data supported the influence of isolation by geographical distance, recent vicariance events, and demographic expansions apparently related to Pleistocene and Holocene forest dynamics.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 73–84.     

Long‐term transportation,by road and air,of chub mackerel (Scomber japonicus) and atlantic bonito (Sarda sarda)

During the second semester of 2009, three trips were made from Olhão (Southern Portugal) to Stralsund (Northern Germany) carrying 2.122 animals, which included multiple teleosts, elasmobranchs and invertebrates. This group included scombrids, such as 1.869 Scomber japonicus and 9 Sarda sarda, which are notoriously difficult to transport. However, multiple adaptations to transport regimes adopted regularly have allowed the authors to successfully move these animals by road and air over a total of up to 25 hr. Such adaptations included maintaining oxygen saturation rates at approximately 200%, and also the constant addition of AmQuel^®, sodium bicarbonate, and sodium carbonate. Different formulations were used during the three trips, with the best results corresponding to 20/30/30 ppm of the three aforementioned chemicals, respectively. The authors suggest, however, that a modified formula of 20/40/40 ppm will allow for an even more stable pH on future trips. Zoo Biol 30:459–472, 2010. © 2010 Wiley‐Liss, Inc.     

Comparative phylogeography of Atlantic bluefin tuna and swordfish: the combined effects of vicariance, secondary contact, introgression, and population expansion on the regional phylogenies of two highly migratory pelagic fishes

Comparative phylogeography has revealed remarkable patterns of concordance in the maternal phylogenies of many species. The phylogeography and historical demography of the mitochondrial control region I for 607 Atlantic bluefin tuna (Thunnus thynnus) and 275 swordfish (Xiphias gladius) were analyzed to clarify the complex phylogenetic signals in the North Atlantic-Mediterranean region where they are sympatric. Atlantic bluefin tuna mtDNA is polyphyletic, and includes rare sequences sister to Pacific bluefin tuna (Thunnus orientalis) and introgressed albacore (Thunnus alalunga) sequences. There is no geographic partitioning between Atlantic and Mediterranean samples of Atlantic bluefin tuna (Phi(ST)=0.002). In contrast, Atlantic and Mediterranean swordfish are differentiated (Phi(ST)=0.091) due to the combined effects of vicariance, secondary contact, and dissimilar regional demographic histories. Mediterranean swordfish has substantially less variation, and a more recent history (tau=2.42) than that of Atlantic swordfish (tau=7.02). In spite of the discordant phylogenetic and phylogeographic signals, the demographic history of Atlantic swordfish and Atlantic bluefin tuna (tau=7.51) suggests concordance in the timeline of population expansion. Possible scenarios of cladogenesis, expansion, and contraction, influenced by glacial cycles during the Pleistocene, are formulated.     

Montane refugia isolation and plateau population expansion: Phylogeography of Sino-Himalayan endemic Spenceria ramalana (Rosaceae)

Climate oscillations and landscape heterogeneity make the historical population processes of organisms of the Qinghai–Tibet Plateau quite complex. We used chloroplast DNA sequences and amplified fragment length polymorphism (AFLPs) to study the phylogeography and genetic variation of the Qinghai–Tibet Plateau endemic Spenceria ramalana Trimen. The parsimony network contains two star-like evolutionary units, nested into two nested clades, coinciding with geographic population groups indicated by samova software. The central group shows genetic homogeneity, indicating a bottleneck or founder effect of population dynamics. Margin groups contribute more haplotype diversity (C_T), indicating potential refugia. Several expansion events during several interglacial periods were detected. The latest expansion of the eastern population contributed to the modern population structure of the central area. Four AFLP clusters were detected. Higher C_T, a relict pattern of haplotype distribution and DW values of marginal populations, confirmed the location of potential montane refugia. Relict distribution pattern, nested clade phylogeographic analysis conclusions, and restricted gene flow detected by AFLPs indicated an isolation pattern of the populations of montane areas. Heterogeneity of landscape of the western and southern montane areas could be the main barrier for gene flow. Moisture conditions and ecotype transformation might help to build the phylogeographic pattern of this species. Neither chloroplast DNA markers nor AFLP was able to differentiate the two varieties of this species.     

Dispersal vs. vicariance in the Mediterranean: historical biogeography of the Palearctic Pachydeminae (Coleoptera, Scarabaeoidea)

Aim The geological evolution of the Mediterranean region is largely the result of the Tertiary collision of the African and Eurasian Plates, but also a mosaic of migrating island arcs, fragmenting tectonic belts, and extending back‐arc basins. Such complex paleogeography has resulted in a ‘reticulate’ biogeographical history, in which Mediterranean biotas repeatedly fragmented and merged as dispersal barriers appeared and disappeared through time. In this study, dispersal‐vicariance analysis (DIVA) is used to assess the relative role played by dispersal and vicariance in shaping distribution patterns in the beetle subfamily Pachydeminae Reitter, 1902 (Scarabaeoidea), an example of east–west Mediterranean disjunction. Location The Mediterranean region, including North Africa, the western Mediterranean, Balkans–Anatolia, Middle East, Caucasus, the Iranian Plateau, and Central Asia. Methods A phylogenetic hypothesis of the Palearctic genera of Pachydeminae in conjunction with distributional data was analysed using DIVA. This method reconstructs the ancestral distribution in a given phylogeny based on the vicariance model, while allowing dispersal and extinction to occur. Unlike other methods, DIVA does not enforce area relationships to conform to a hierarchical ‘area cladogram’, so it can be used to reconstruct ‘reticulate’ biogeographical scenarios. Results Optimal reconstructions, requiring 23 dispersal events, suggest that the ancestor of Pachydeminae was originally present in the south‐east Mediterranean region. Basal splitting within the subfamily was caused by vicariance events related to the late Tertiary collision of the African microplates Apulia and Arabia with Eurasia, and the resultant arise of successive dispersal barriers (e.g. the Red Sea, the Zagros Mountains). Subsequent diversification in Pachydeminae involved multiple speciation events within the Middle East and Iran–Afghanistan regions, which gave rise to the least speciose genera of Pachydeminae (e.g. Otoclinius Brenske, 1896). Finally, the presence of Pachydeminae in the western Mediterranean region seems to be the result of a recent dispersal event. The ancestor of the Iberian genera Ceramida Baraud, 1987 and Elaphocera Gené, 1836 probably dispersed from the Middle East to the Iberian Peninsula across North Africa and the Gibraltar Strait during the ‘Messinian salinity crisis’ at the end of the Miocene. Main conclusions Although the basal diversification of Pachydeminae around the Mediterranean appears to be related to vicariance events linked to the geological formation of the Mediterranean Basin, dispersal has also played a very important role. Nearly 38% of the speciation events in the phylogeny resulted from dispersal to a new area followed by allopatric speciation between lineages. Relationships between western and eastern Mediterranean disjuncts are usually explained by dispersal through Central Europe. The biogeographical history of the Pachydeminae corroborates other biogeographical studies that consider North Africa to be an alternative dispersal route by which Mediterranean taxa could have achieved circum‐Mediterranean distributions.     

Phylogeography and population dynamics of the white-sided dolphin (Lagenorhynchus acutus) in the North Atlantic

Highly mobile species in the marine environment may be expected to show little differentiation at the population level, but this is often not the case. Instead cryptic population structure is common, and effective conservation will require an understanding of how these patterns evolve. Here we present an assessment from both sides of the North Atlantic of differentiation among populations of a dolphin species that inhabits mainly pelagic waters, the Atlantic white-sided dolphin. We compare eleven putative populations in the western and eastern North Atlantic at mtDNA and microsatellite DNA loci and find reduced nucleotide diversity and signals for historical bottlenecks and post-bottleneck expansions in all regions. We calculate expansion times to have occurred during the early Holocene, following the last glacial maximum (LGM). We find evidence for connectivity among populations from either side of the North Atlantic, and differentiation between putative populations in the far northeast compared with all other areas sampled. Some data suggest the possibility of separate refugia during the LGM explaining this pattern, although ongoing ecological processes may also be a factor. We discuss the implications for developing effective programs of conservation and management in the context of ongoing anthropogenic impact.     

Phylogeography of the Atlanto-Mediterranean sea cucumber Holothuria (Holothuria) mammata: the combined effects of historical processes and current oceanographical pattern

We assessed the genetic structure of populations of the widely distributed sea cucumber Holothuria (Holothuria) mammata Grube, 1840, and investigated the effects of marine barriers to gene flow and historical processes. Several potential genetic breaks were considered, which would separate the Atlantic and Mediterranean basins, the isolated Macaronesian Islands from the other locations analysed, and the Western Mediterranean and Aegean Sea (Eastern Mediterranean). We analysed mitochondrial 16S and COI gene sequences from 177 individuals from four Atlantic locations and four Mediterranean locations. Haplotype diversity was high (H=0.9307 for 16S and 0.9203 for COI), and the haplotypes were closely related (π=0.0058 for 16S and 0.0071 for COI). The lowest genetic diversities were found in the Aegean Sea population. Our results showed that the COI gene was more variable and more useful for the detection of population structure than the 16S gene. The distribution of mtDNA haplotypes, the pairwise F(ST) values and the results of exact tests and amova revealed: (i) a significant genetic break between the population in the Aegean Sea and those in the other locations, as supported by both mitochondrial genes, and (ii) weak differentiation of the Canary and Azores Islands from the other populations; however, the populations from the Macaronesian Islands, Algarve and West Mediterranean could be considered to be a panmictic metapopulation. Isolation by distance was not identified in H. (H.) mammata. Historical events behind the observed findings, together with the current oceanographic patterns, were proposed and discussed as the main factors that determine the population structure and genetic signature of H. (H.) mammata.     

Phylogeography and phylogeny of the genus Acanthonyx (Decapoda,Epialtidae) in the north‐east Atlantic and Mediterranean

The genus Acanthonyx Latreille, 1828 (Majoidea, Epialtidae) contains 17 or 18 known species, depending on competing taxonomic views, that are widely distributed across the world. Morphologically, most species look superficially alike, and therefore, similar taxonomic concepts have been described under different names. Consequently, there is today a considerable list of synonyms, which further complicates the taxonomy of the genus Acanthonyx. In this study, we conducted a phylogeographical and phylogenetic analysis of populations of the genus Acanthonyx in the NE Atlantic, Mediterranean and Macaronesia using the mitochondrial cytochrome oxidase subunit I (COI) and the nuclear 28S rRNA loci. Our phylogenetic and phylogeographical results revealed that Acanthonyx lunulatus sensu lato is a complex of three distinct lineages: one corresponding to the previously described Acanthonyx brevifrons, another to A. lunulatus sensu stricto and a third to a yet undescribed group. Whereas our results confirms that A. brevifrons deserves the status of a species, as it can be easily distinguishable from A. lunulatus by a few morphological traits, we could not find any such traits suitable for the discrimination between A. lunulatus sensu stricto and the third lineage. Furthermore, the degree of COI divergence between this lineage and A. lunulatus is below average levels for Decapoda species. Yet, no shared haplotypes have been detected between them. The differences found in the nuclear gene (indels), together with the sympatric occurrence of the two forms, prompt for a more detailed analysis of this group. Overall, the results show that significant genetic differentiation between specimens with similar morphology occurs in the Epialtidae, thus reinforcing the importance of combining morphological and genetic tools to fully resolve the taxonomy of these decapods.     

Intraspecific phylogeographic genomics from multiple complete mtDNA genomes in Atlantic cod (Gadus morhua): origins of the "codmother," transatlantic vicariance and midglacial population expansion

On the basis of multiple complete mitochondrial DNA genome sequences, we describe the temporal phylogeography of Atlantic cod (Gadus morhua), a lineage that has undergone a complex pattern of vicariant evolution, postglacial demographic shifts, and historic sharp population declines due to fishing and/or environmental shifts. Each of 32 fish from four spawning aggregations from the northwest Atlantic and Norway has a unique mtDNA sequence, which differs by 6-60 substitutions. Phylogenetic analysis identifies six major haplogroups that range in age from 37 to 75 KYA. The widespread haplotype identified by previous single-locus analyses at the center of a "star phylogeny" is shown to be a paraphyletic assemblage of genome lineages. The coalescent that includes all cod occurs 162 KYA. The most basal clade comprises two fish from the western Atlantic. The most recent superclade that includes all fish examined from Norway, and which includes 84% of all fish examined, dates to 128 KYA at the Sangamon/Würm interglacial, when ocean depths on continental shelves would have favored transcontinental movement. The pairwise mismatch distribution dates population expansion of this superclade to the middle of the Wisconsinan/Weichsel glaciation 59 KYA, rather than to a postglacial emergence from a marine refugium 12 KYA, or to more recent historic events. We discuss alternative scenarios for the expansion and distribution of the descendants of the "codmother" in the North Atlantic. Mitochondrial phylogenomic analyses generate highly resolved trees that enable fine-scale tests of temporal hypotheses with an accuracy not possible with single-locus methods.     

Phylogeography of the common vampire bat (Desmodus rotundus): Marked population structure, Neotropical Pleistocene vicariance and incongruence between nuclear and mtDNA markers

Background  

The common vampire bat Desmodus rotundus is an excellent model organism for studying ecological vicariance in the Neotropics due to its broad geographic range and its preference for forested areas as roosting sites. With the objective of testing for Pleistocene ecological vicariance, we sequenced a mitocondrial DNA (mtDNA) marker and two nuclear markers (RAG2 and DRB) to try to understand how Pleistocene glaciations affected the distribution of intraspecific lineages in this bat.     

Phylogeography of the Temminck’s Stint (Calidris temminckii): historical vicariance but little present genetic structure in a regionally endangered Palearctic wader

Aim We study the population differentiation and phylogeography of the Temminck’s Stint (Calidris temminckii). Specifically, we seek signs of past and present population size changes and dispersal events and evaluate management and conservation unit status of the populations. We also study the possibility of introgression as the origin of two mitochondrial DNA (mtDNA) lineages found and estimate the divergence time of the lineages. Location Northern Eurasia. Methods We analysed 583 bp of mtDNA control region domains I and II and 11 microsatellite loci from 13 localities throughout the breeding range. In addition, we used mitochondrial cytochrome c oxidase subunit I (COI), a barcoding gene, to search for signs of introgression. Results More population differentiation was found from microsatellites than from mtDNA, although differentiation was weak in both markers. Signs of past population growth were observed, in addition to more recent decline in some areas. Both control region and COI sequences revealed two maternal lineages coexisting in Fennoscandia and in north‐west Siberia. No signs of introgression were detected. Lineage divergence time was estimated to have occurred during the glacial periods of Pleistocene. Main conclusions Slight differences in mtDNA and microsatellite differentiation and diversity may reflect different features – such as the mutation rate and effective population size – of the markers used, or female‐biased dispersal pattern and high male site‐fidelity of the species. The coexistence of the two mitochondrial lineages is most likely a consequence of post‐glacial mixing of two refugial Pleistocene populations. Based on genetic information alone, global conservation concerns are not imminent. However, fast decline of a marginal Bothnian Bay population and the smallness and remoteness of a Central Yakutian population warrant conservation actions.     

Phylogeography of the freshwater crayfish Cherax destructor Clark (Parastacidae) in inland Australia: historical fragmentation and recent range expansion

The yabby, Cherax destructor Clark, is the most widespread species in the most widespread genus of Australian freshwater crayfish. It has a distribution that spans several distinct drainage basins and biogeographical regions within semiarid and arid inland Australia. Here we report a study designed to investigate patterns of genetic variation within the species and hypotheses put forward to account for its extensive distribution using DNA sequences from the mitochondrial 16S rRNA gene region. Results of phylogenetic analyses contradicted previous allozyme data and revealed relatively deep phylogenetic structure in the form of three geographically correlated clades. The degree of genetic divergences between clades (8–15 bp) contrasted with the relatively limited haplotype diversity within clades (1–3 bp). Network-based analyses confirmed these results and revealed genetic structure on both larger and more restricted geographical scales. Nevertheless some haplotypes and 1-step clades had large distributions, some of which crossed boundaries between river basins and aquatic biogeographical regions. Thus both older and more recent historical processes, including fragmentation on a larger geographical scale and more recent range expansion on a local scale, appear to be responsible for the observed pattern of genetic variation within C. destructor . These results support elements of alternative hypotheses previously put forward to account for the evolutionary history of C. destructor and the origin of its large distribution.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 539–550.     

Range expansion by invasion: genetic characterization of invasion of the greenside darter (Etheostoma blennioides) at the northern edge of its distribution

Species introductions in freshwater ecosystems are often complex processes, yet an understanding of the nature of the introduction can inform management and conservation actions. The greenside darter (Etheostoma blennioides), until recently a species of special concern, expanded its Canadian range and is now common and widespread in the Grand River watershed (GRW). This is despite there being no evidence of greenside darter in the GRW prior to 1990. The goal of this study was to genetically characterize the GRW greenside darter introduction. Greenside darter were sampled in the GRW, the three known native watersheds in Canada, and one site from Ohio. We measured genetic diversity and population structure, and tested for population bottlenecks using eight microsatellite loci. Genotype assignment was used to identify possible introduction sources. Populations in the GRW showed similar genetic diversity to native watershed populations with no evidence for recent or historical population bottlenecks. Genotype assignment showed that one of the Canadian watersheds and the Ohio site were not potential sources of the GRW greenside darter, whereas the Thames River watershed was the most likely source. Substantial population genetic structure exists among the sample sites in the GRW. Clearly, the current widespread and abundant distribution of the greenside darter in the GRW is not the result of recent expansion of an existing native population, but rather multiple introductions into at least three sites in the GRW, followed by rapid population growth. Although the GRW E. blennioides is introduced, it harbours considerable genetic diversity and represents an important northern range extension for this species.     

Genetic divergence and isolation by distance in the West Atlantic sea anemone Actinia bermudensis (McMurrich, 1889)

The sea anemone Actinia bermudensis [Proc. Acad. Nat. Sci. Philadelphia 41 (1889) 102] has a widespread distribution over the Western Atlantic, where it has been reported from Bermuda and Bahamas to southern Brazil. Nonetheless, reproductive features of this species, which broods asexually produced offspring, suggest that it has low dispersal capabilities. This raises a question about the specific status of the Brazilian population of A. bermudensis. The aim of this paper was to evaluate the taxonomic status of the Brazilian population, by genetically comparing it with the type locality (Bermuda) population. For this, we used allozyme electrophoresis to analyze samples from both populations. Thirteen enzyme systems (16 loci) yielded interpretable results for all populations. A high differentiation was encountered for the Western Atlantic A. bermudensis population (F_ST=0.434), partially due to the presence of a diagnostic locus (LAP) between Bermuda and Brazilian populations. This result could be taken as an indication that the Brazilian population should be elevated to a specific status. We do expect some genetic differentiation, however, between natural populations separated over wide geographic distances. Finally, since Mantel and regression analyses did not rule out the isolation by distance model, we have conservatively decided to maintain the specific status of A. bermudensis from Brazil.